Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays

Viruses

JuanC. de la Torre¹  Beatrice Cubitt¹  Masaharu Iwasaki¹  Yíngyún Caì²  PeterB. Jahrling²  LisaEvans DeWald²  ElenaN. Postnikova²  Shuīqìng Yú²  JensH. Kuhn²  Laura Bollinger²  GustavoF. Palacios³  SheliR. Radoshitzky³  BrettF. Beitzel³ 

[1] Department of Immunology and Microbial Science, The Scripps Research Institute (TSRI), 10550 North Torrey Pines Rd., La Jolla, CA 92037, USA;Integrated Research Facility at Fort Detrick (IRF-Frederick), National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), B-8200 Research Plaza, Fort Detrick, Frederick, MD 21702, USA;United States Army Medical Research Institute of Infectious Diseases (USAMRIID), 1425 Porter Street, Fort Detrick, Frederick, MD 21702, USA;

关键词: Arenaviridae;    arenavirus;    biosafety level 4;    BSL-4;    drug screening;    Lassa fever;    Lassa virus;    LASV;    mammarenavirus;    reverse genetics;   

DOI  :  10.3390/v10110655

来源: DOAJ

【 摘 要 】

Lassa virus (LASV), a mammarenavirus, infects an estimated 100,000⁻300,000 individuals yearly in western Africa and frequently causes lethal disease. Currently, no LASV-specific antivirals or vaccines are commercially available for prevention or treatment of Lassa fever, the disease caused by LASV. The development of medical countermeasure screening platforms is a crucial step to yield licensable products. Using reverse genetics, we generated a recombinant wild-type LASV (rLASV-WT) and a modified version thereof encoding a cleavable green fluorescent protein (GFP) as a reporter for rapid and quantitative detection of infection (rLASV-GFP). Both rLASV-WT and wild-type LASV exhibited similar growth kinetics in cultured cells, whereas growth of rLASV-GFP was slightly impaired. GFP reporter expression by rLASV-GFP remained stable over several serial passages in Vero cells. Using two well-characterized broad-spectrum antivirals known to inhibit LASV infection, favipiravir and ribavirin, we demonstrate that rLASV-GFP is a suitable screening tool for the identification of LASV infection inhibitors. Building on these findings, we established a rLASV-GFP-based high-throughput drug discovery screen and an rLASV-GFP-based antibody neutralization assay. Both platforms, now available as a standard tool at the IRF-Frederick (an international resource), will accelerate anti-LASV medical countermeasure discovery and reduce costs of antiviral screens in maximum containment laboratories.

【 授权许可】

Unknown